分子生物学
聚合酶链反应
生物
寡核苷酸
免疫球蛋白轻链
热启动PCR
克隆(Java方法)
免疫球蛋白重链
基因
底漆(化妆品)
抗体
克隆(编程)
DNA
遗传学
化学
套式聚合酶链反应
有机化学
程序设计语言
计算机科学
作者
James W. Larrick,Lena Danielsson,Carol A. Brenner,Magnus Abrahamson,Kirk E. Fry,Carl Borrebaeck
标识
DOI:10.1016/s0006-291x(89)80138-x
摘要
A general method to directly obtain the DNA sequence of the variable regions of any immunoglobulin chain using a mixture of oligomer primers and the polymerase chain reaction (PCR) is described. Mixed oligonucleotide primers corresponding to the 5′ signal peptide and a conserved 3′ constant region primer were used for enzymatic amplification of each of the heavy and light chain variable regions of a human hybridoma producing a monoclonal antibody recognizing an epitope of gp120 of the human immunodeficiency virus 1. The amplified DNA segments were cloned and the sequence was determined for the heavy chain variable region. This method will greatly facilitate structural and functional studies of immunoglobulins by reducing the effort to clone and sequence the members of the immunoglobulin as well as other multigene families.
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